Title

Detangling the Web: Isolation of the MaSp1 Promoter from the Black Widow spider

Poster Number

13

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Abstract/Artist Statement

The long-term objective of this research experiment is to understand how the expression of silk genes is regulated in black widow spiders. Syuthetic silk production will be possible in the near future but understanding how a spider expresses these genes is one of the most critical steps. In this research proposal, we attempted to clone the promoter region of the fibroin MaSp 1 from the black widow spider. Different regions of the MaSpl promoter were amplified using polymerasechain reaction using different primers that were designed to amplify the 5' -flanking QNA region of the MaSpl gene. We were able to successfully amplify promoter regions sized at 300 bp, 1000 bp-and 1500 bp. After amplification of the different promoter regions from genomic DNA, we inserted the fragments into the cloning vector pBAD-TOPO. Acquisition of the promoter fragments were confirmed by restriction digestion and agarose gel electrophoresis. Our future goal is to insert these fragments into a promoterless reporter plasmid and measure the transcriptional activity of these DNA regions in insect cells.

Location

Wendell Phillips Center, 1st floor hallways

Start Date

3-5-2008 1:00 PM

End Date

3-5-2008 3:00 PM

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May 3rd, 1:00 PM May 3rd, 3:00 PM

Detangling the Web: Isolation of the MaSp1 Promoter from the Black Widow spider

Wendell Phillips Center, 1st floor hallways

The long-term objective of this research experiment is to understand how the expression of silk genes is regulated in black widow spiders. Syuthetic silk production will be possible in the near future but understanding how a spider expresses these genes is one of the most critical steps. In this research proposal, we attempted to clone the promoter region of the fibroin MaSp 1 from the black widow spider. Different regions of the MaSpl promoter were amplified using polymerasechain reaction using different primers that were designed to amplify the 5' -flanking QNA region of the MaSpl gene. We were able to successfully amplify promoter regions sized at 300 bp, 1000 bp-and 1500 bp. After amplification of the different promoter regions from genomic DNA, we inserted the fragments into the cloning vector pBAD-TOPO. Acquisition of the promoter fragments were confirmed by restriction digestion and agarose gel electrophoresis. Our future goal is to insert these fragments into a promoterless reporter plasmid and measure the transcriptional activity of these DNA regions in insect cells.